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Background <p>Putrefaction is the microbial metabolism/degradation of autolysis end products like amino acids. The aim of this study was to identify and investigate amino acids and their bacterial degradation products/metabolites by LC-HRMS as possible markers of putrefaction.</p> Methods <p>Pig eyes (<i>n</i> = 60) were stored in conical centrifuge tubes and petri dishes within a climate chamber at 25°C, 65% humidity over four days. Each day, eyes were macroscopically examined and vitreous humour was drawn from up to six eyes. Twenty-one amino acids/metabolites (lysine, histidine, arginine, <i>N</i><sup><i>2</i></sup>-acetyllysine, <i>N</i><sup><i>2</i></sup>-acetylarginine, <i>N</i><sup><i>6</i></sup>-acetyllysine, <i>N</i>-acetylcadaverine, tyrosine, <i>N</i>-acetylagmatine, tyramine, phenylalanine, <i>N</i><sup><i>2,6</i></sup>-diacetyllysine, 2-phenylethylamine, tryptophan, kynurenic acid, <i>N-</i>acetyltyrosine, <i>N</i>-acetyltyramine, <i>N</i>-acetylphenylalanine, <i>N</i>-acetyltryptophan, <i>N</i>-acetyl-2-phenylethylamine, <i>N</i>-acetyltryptamine<b>)</b> were monitored by LC-HRMS.</p> Results <p>Opacification was observed and eyes stored on petri dishes dried out faster and turned dark compared to eyes stored in centrifuge tubes. Amino acids were universally present and showed a significant increase by up to 46-fold from day 0 to 4, except for arginine (no trend observed). Amino acid metabolites were first detected on day 2 with exception of <i>N</i><sup><i>2</i></sup>-acetyllysine, <i>N</i><sup><i>2</i></sup>-acetylarginine, and <i>N</i><sup><i>6</i></sup>-acetyllysine (detected on day 0). All analytes showed a marked increase by days 3 and 4, especially kynurenic acid and <i>N</i>-acetylated compounds. Among the six eyes of each storage condition/day high variability was observed, between centrifuge tube and petri dish storage few variability was found.</p> Conclusion <p>LC-HRMS analysis of amino acids and their metabolites in vitreous humour could be a promising tool to evaluate the putrefaction status. However, further studies are required to better understand inter-individual metabolic differences and influencing environmental factors.</p>

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Amino acids and their metabolites as potential biochemical markers in postmortem vitreous humour

  • 期刊论文

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  • 发表日期：

    2025年7月2日

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  • Laura Franke,
  • Hannah Ihle,
  • Kristina Rieger,
  • Viviane Stammer,
  • Senta Niederegger,
  • Dirk K. Wissenbach,
  • Frank T. Peters,
  • Gita Mall

摘要

Background

Putrefaction is the microbial metabolism/degradation of autolysis end products like amino acids. The aim of this study was to identify and investigate amino acids and their bacterial degradation products/metabolites by LC-HRMS as possible markers of putrefaction.

Methods

Pig eyes (n = 60) were stored in conical centrifuge tubes and petri dishes within a climate chamber at 25°C, 65% humidity over four days. Each day, eyes were macroscopically examined and vitreous humour was drawn from up to six eyes. Twenty-one amino acids/metabolites (lysine, histidine, arginine, N2-acetyllysine, N2-acetylarginine, N6-acetyllysine, N-acetylcadaverine, tyrosine, N-acetylagmatine, tyramine, phenylalanine, N2,6-diacetyllysine, 2-phenylethylamine, tryptophan, kynurenic acid, N-acetyltyrosine, N-acetyltyramine, N-acetylphenylalanine, N-acetyltryptophan, N-acetyl-2-phenylethylamine, N-acetyltryptamine) were monitored by LC-HRMS.

Results

Opacification was observed and eyes stored on petri dishes dried out faster and turned dark compared to eyes stored in centrifuge tubes. Amino acids were universally present and showed a significant increase by up to 46-fold from day 0 to 4, except for arginine (no trend observed). Amino acid metabolites were first detected on day 2 with exception of N2-acetyllysine, N2-acetylarginine, and N6-acetyllysine (detected on day 0). All analytes showed a marked increase by days 3 and 4, especially kynurenic acid and N-acetylated compounds. Among the six eyes of each storage condition/day high variability was observed, between centrifuge tube and petri dish storage few variability was found.

Conclusion

LC-HRMS analysis of amino acids and their metabolites in vitreous humour could be a promising tool to evaluate the putrefaction status. However, further studies are required to better understand inter-individual metabolic differences and influencing environmental factors.